Regulation of EMT During Avian Heart Valve Formation

禽类心脏瓣膜形成过程中 EMT 的调节

• 批准号: 7598984
• 负责人: RAYMOND Bruce RUNYAN
• 金额: $ 37.75万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7598984
• 关键词: ACVR1 gene; Adult; Antibodies; Antisense Oligonucleotides; Biological Assay; Biology; Birds; Cardiac; Cell Death; Cell Separation; Cell physiology; Cells; Collagen; Complex; Cytoskeleton; Data; Development; Dimensions; Dissociation; ERBB2 gene; Elements; Embryo; Embryonic Heart; Endoglin; Endothelial Cells; Epithelial; Epithelium; Extracellular Matrix; Family; Fibrosis; Gel; Gene Expression; Genes; Growth Factor; Heart; Heart Valves; Homeobox; Hyaluronan; Islets of Langerhans; Kidney; Laboratories; Laboratory Study; Lateral; Ligands; Light; Mediating; Mediator of activation protein; Mesenchymal; Modeling; Mus; Neoplasm Metastasis; Neural Crest; Neural Crest Cell; Neural tube; Neuregulins; Outcome; Pathologic Processes; Peptide Hydrolases; Phenotype; Population; Process; Production; Protein Isoforms; Proteins; Regulation; Role; Shapes; Signal Transduction; Site; Somites; Staging; Surface; System; TGF-beta type I receptor; TGFB1 gene; TGFB3 gene; Time; Tissues; Transforming Growth Factor Beta 2; Tricuspid valve structure; Variant; Vascular Endothelial Growth Factors; Vertebrates; Work; Wound Healing; Zebrafish; Zinc Fingers; cadherin 5; cardiogenesis; cell motility; cell transformation; cell type; congenital heart disorder; experimental analysis; gastrulation; migration; novel; progenitor; receptor; receptor expression; research study; response; slug; transcription factor; two-dimensional

DESCRIPTION (provided by applicant): During cardiac development, valve progenitors first arise in the atrioventricular canal as a result of a TGF¿-mediated epithelial-mesenchymal cell transformation (EMT). Prior studies showed that this EMT can be divided into two stages, an activation stage and an invasion stage. Stage 1 is mediated, in part, by transforming growth factor beta 2 (TGF¿2) and stage 2 is mediated, in part, by TGF¿3. Previous work showed that EMT can be disrupted by the inhibition of 5 different TGF¿ receptors, TGF¿ receptor II, TGF¿ receptor III, Endoglin, ALK2 and ALK5. It is our hypothesis that there are two more discrete signaling complexes mediating TGF¿-regulated EMT. To continue the analysis of TGF¿-mediated regulation of normal heart development, the first aim will examine the correlation between specific TGF¿ isoforms and receptors in mediating EMT regulation. Microarray experiments will be performed to assess the effect loss of specific TGF¿ isoforms on altered gene expression in tissues prior to EMT, during EMT and in mesenchymal cells after EMT. Selected markers among the TGF¿-regulated genes will then be used to assess whether gene expression is altered as each of the specific receptors is disrupted. Correlations between these data will point to specific receptor and ligand signaling complexes that mediate EMT. These putative complexes will be confirmed at the protein level. Data from other labs has implicated a variety of signal transduction mechanisms and intracellular regulators in cardiac EMT including ErbB2, NF-1, VEGF and NFATc1. In the second aim, experiments will be undertaken to determine whether disruption of TGF¿ signaling alters any of the other candidate mechanisms or whether loss of these molecules alters TGF¿ ligand or receptor expression. It was observed in the neural crest system that forced expression of Sox 8, 9 or 10 in the neural tube would cause an EMT and produce ectopic neural crest cells. We conjecture that this is analogous to the cell invasion step in the heart and that it requires some TGF¿ activity. In the third aim, experiments will be undertaken to determine whether exogenous expression of Sox genes will force EMT and whether Sox gene expression is regulated by TGF¿ signal transduction or regulates TGF¿-mediated responses. Together these experiments will shed new light on the mechanisms of EMT regulation and aid in an understanding of congenital heart disease.

描述（由申请人提供）：在心脏发育期间，由于TGF β介导的上皮-间充质细胞转化（EMT），瓣膜祖细胞首先在房室管中出现。以往的研究表明，这种EMT可以分为两个阶段，激活阶段和入侵阶段。阶段1部分由转化生长因子β 2（TGF β 2）介导，阶段2部分由TGF β 3介导。以前的工作表明，EMT可以通过抑制5种不同的TGF β受体，TGF β受体II，TGF β受体III，内皮糖蛋白，ALK 2和ALK 5来破坏。我们的假设是，有两个以上的离散信号复合物介导TGF β调节EMT。为了继续分析TGF β介导的正常心脏发育调节，第一个目的将检查特定TGF β亚型和受体在介导EMT调节中的相关性。将进行微阵列实验，以评估特定TGF β亚型对EMT前、EMT期间和EMT后间充质细胞中组织中基因表达改变的影响损失。然后，在TGF β-调节基因中选择的标记物将用于评估基因表达是否随着每个特定受体的破坏而改变。这些数据之间的相关性将指向介导EMT的特定受体和配体信号传导复合物。这些假定的复合物将在蛋白质水平上得到证实。来自其他实验室的数据表明，心脏EMT中存在多种信号转导机制和细胞内调节因子，包括ErbB 2、NF-1、VEGF和NFATc 1。在第二个目标中，将进行实验以确定TGF β信号传导的中断是否改变任何其他候选机制，或者这些分子的丢失是否改变TGF β配体或受体表达。在神经嵴系统中观察到，在神经管中Sox 8、9或10的强制表达将引起EMT并产生异位神经嵴细胞。我们推测这类似于心脏中的细胞侵入步骤，并且它需要一些TGF？活性。在第三个目标中，将进行实验以确定Sox基因的外源性表达是否会迫使EMT，以及Sox基因的表达是否受TGF β信号转导或调节TGF β介导的反应的调节。这些实验将为EMT调节机制提供新的线索，并有助于了解先天性心脏病。